We propose to develop improved procedures for the analysis of biomedical samples which utilize the high sensitivity of laser fluorimetry, coupled with high pressure liquid chromatography, as a means of realizing greater specificity, lower limits of detection, shorter analysis times, and simpler experimental protocols. We will continue our work in the development of laser fluorescence immunoassay methods, in which the detection of insulin is used as a prototype system. Our goal is to develop viable, generally applicable alternatives to radioimmunoassay. One route proposed will replace the radioactive tag with a label whose unique fluorescence properties afford a means of selectively rejecting the background luminescence which generally hampers the practical application of fluorescence immunoassay methods. Another approach we will pursue combines laser fluorimetry with enzymatic amplification to increase the sensitivity and rapidity of enzyme-linked immunoassay. In addition, we propose to develop sensitive laser fluorimetric methods for the analysis of biogenic amines using fluorescence derivatization techniques. We propose specific applications to analyses for opioid peptides, such as enkephalins, and the slow-reacting substances of anaphylaxis (SRS-A). Applications to the detection of amino acid residues in protein sequencing analyses also are proposed.